title: SYBR®Green qPCR Salmonella detection system allowing discrimination at the genus, species and subspecies levels.
abstract: In this work, a three-level Salmonella detection system based on a combination of seven SYBR®Green qPCR was developed. This detection system discriminates Salmonella at the genus, species and subspecies levels using a single 96-well plate. The SYBR®Green qPCR assays target the invA, rpoD, iroB and safC genes, as well as the STM0296 locus, putatively coding for a cytoplasmic protein. This study includes the design of primer pairs, in silico and in situ selectivity, sensitivity, repeatability and reproducibility evaluations of the seven SYBR®Green qPCR assays. Each detection level displayed a selectivity of 100 %. This combinatory SYBR®Green qPCR system was also compared with three commercially available Salmonella qPCR detection kits. This comparison highlighted the importance of using a multi-gene detection system to be able to detect every target strain, even those with deletion or mutation of important genes.
authors.affiliation: Scientific Institute of Public Health, J. Wytsmanstraat 14, 1050, Brussels, Belgium.
authors.forename: Elodie
authors.initials: E
authors.lastname: Barbau-Piednoir
authors.forename: Sophie
authors.initials: S
authors.lastname: Bertrand
authors.forename: Jacques
authors.initials: J
authors.lastname: Mahillon
authors.forename: Nancy H
authors.initials: NH
authors.lastname: Roosens
authors.forename: Nadine
authors.initials: N
authors.lastname: Botteldoorn
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D001431:Bacteriological Techniques / Q000379:methods*; D052160:Benzothiazoles; D017931:DNA Primers / Q000235:genetics; D003959:Diamines; D005456:Fluorescent Dyes / Q000378:metabolism*; D009930:Organic Chemicals / Q000378:metabolism*; D011804:Quinolines; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012475:Salmonella / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods
chemical_list: D001426:Bacterial Proteins; D052160:Benzothiazoles; D017931:DNA Primers; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I